FIG 4.

Stably transduced DC2.4 cells expressing γ34.5 are sensitive to starvation and accumulate long-lived proteins. (A) Annexin V staining of DC2.4 cells stably expressing γ34.5 or the ΔBBD mutant or control cells. Cells were untreated or starved and analyzed by flow cytometry, and mean fluorescence intensity was recorded. Graph shows the mean from two experiments, each consisting of at least 2 biological duplicates, and error bars indicate standard deviations. ***, P < 0.001 (t test). (B) Representative Western blot (of three experiments) of p62 or α-tubulin on cell lysates of control cells or cells expressing γ34.5 or the ΔBBD mutant. Cells were untreated or incubated with 100 µM bafilomycin for 6 h. (C) Quantification of a representative Western blot showing band density of p62 normalized to α-tubulin band density.